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Identification of methylation profile of HOX genes in extrahepatic cholangiocarcinoma

World Journal of Gastroenterology 2011年第29期17卷 3407-3419页

作者： Yi Shu Bing Wang Ji Wang Jian-Ming Wang Sheng-Quan Zou General Surgery Department of Tongji Hospital Tongji Medical College of Huazhong University of Science and Technology1095 Jiefang AvenueWuhan 430030Hubei ProvinceChina

AIM:To identify methylation profile and novel tumor marker of extrahepatic cholangiocarcinoma(CCA)with high throughout ***:Differential methylation profile was compared between normal bile duct epithelial cell lines and CCA cell lines by methyl-DNA immunoprecipitation (MeDIP)***-polymerase chain reaction (BSP)was performed to identify the methylated allels of target *** of target genes was investigated before and after the treatment with DNA demethylating *** of candidate genes was also evaluated by immunofluorescence in 30 specimens of CCA tissues and 9 normal bile duct ***:Methylation profile of CCA was identified with MeDIP microarray in the respects of different gene functions and signaling ***,97 genes with hypermethylated CpG islands in the promoter region were homeobox *** top 5 hypermethylated homeobox genes validated by BSP were HOXA2(94.29%),HOXA5(95.38%),HOXA11 (91.67%),HOXB4(90.56%)and HOXD13(94.38%).Expression of these genes was reactivated with 5’-aza2’-*** expression differences were found between normal bile duct and extrahepatic CCA tissues(66.67%-100%vs 3.33%-10%).CONCLUSION:HOXA2,HOXA5,HOXA11,HOXB4 and HOXD13 may work as differential epigenetic biomarkers between malignant and benign biliary tissues.

关键词： DNA methylation Epigenetic promoter microarray Cholangiocarcinoma

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Identification and validation of novel C/EBPp-regulated genes in preadipocyte proliferation

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Chinese Medical Journal 2010年第9期123卷 1190-1194页

作者： LIU Mei HUANG Hai-yan Key Laboratory of Molecular Medicine the Ministry of Education Department of Biochemistry and Molecular Biology Fudan University Shanghai Medical College Shanghai 200032China

Background CCAAT/enhancer-binding protein β（C/EBPβ） is required for mitotic clonal expansion （MCE） during adipogenesis. It is still unclear how C/EBPp regulates MCE in the earlier differentiation programs of 3T3-L1 preadipocytes. The purpose of this paper was to understand why C/EBPβ is required for preadipocyte proliferation, and identify new target genes of C/EBPP with chromatin immunoprecipitation （ChlP）-on-chip. Methods Postconfluent growth-arrested 3T3-L1 preadipocytes were induced to differentiation using a standard differentiation protocol. Chip was performed at 20 hours after induction with specific anti-C/EBPβ antibodies. The precipitated DNA was amplified, labeled and hybridized with a mouse promoter microarray. Compared with the control in which the ChIP experiment was performed with non-specific antibody, only the genes with a signal increasing more than 2 fold were considered as candidate *** A total of 110 candidate genes were identified. BTG3 associated nuclear protein （SMAR1, Banp） and tripartite motif-containing 35 （Hls5, trim35） were two target genes among the 110 candidate genes which are involved in cell cycle regulation; the binding of C/EBP3 to the promoter of banp and trim35 was verified by ChlP-PCR. Conclusion C/EBPβ may regulate preadipocyte proliferation through activation of banp and trim35.

关键词： CCAAT/enhancer binding protein β chromatin immunoprecipitation promoter microarray preadipocyte proliferation

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